Cell Wall Composition as a Marker of the Reprogramming of the Cell Fate on the Example of a Daucus carota (L.) Hypocotyl in Which Somatic Embryogenesis Was Induced.

Changes in the composition of the cell walls are postulated to accompany changes in the cell's fate. We check whether there is a relationship between the presence of selected pectic, arabinogalactan proteins (AGPs), and extensins epitopes and changes in cell reprogramming in order to answer the question of whether they can be markers accompanying changes of cell fate. Selected antibodies were used for spatio-temporal immunolocalization of wall components during the induction of somatic embryogenesis. Based on the obtained results, it can be concluded that (1) the LM6 (pectic), LM2 (AGPs) epitopes are positive markers, but the LM5, LM19 (pectic), JIM8, JIM13 (AGPs) epitopes are negative markers of cells reprogramming to the meristematic/pluripotent state; (2) the LM8 (pectic), JIM8, JIM13, LM2 (AGPs) and JIM11 (extensin) epitopes are positive markers, but LM6 (pectic) epitope is negative marker of cells undergoing detachment; (3) JIM4 (AGPs) is a positive marker, but LM5 (pectic), JIM8, JIM13, LM2 (AGPs) are negative markers for pericycle cells on the xylem pole; (4) LM19, LM20 (pectic), JIM13, LM2 (AGPs) are constitutive wall components, but LM6, LM8 (pectic), JIM4, JIM8, JIM16 (AGPs), JIM11, JIM12 and JIM20 (extensins) are not constitutive wall components; (5) the extensins do not contribute to the cell reprogramming.

Endophytic bacterial communities in peels and pulp of five root vegetables.

Plants contain endophytic bacteria, whose communities both influence plant growth and can be an important source of probiotics. Here we used deep sequencing of a 16S rRNA gene fragment and bacterial cultivation to independently characterize the microbiomes of five plant species from divergent taxonomic orders-potato (Solanum tuberosum), carrot (Daucus sativus), beet (Beta vulgaris), neep (Brassica napus spp. napobrassica), and topinambur (Helianthus tuberosus). We found that both species richness and diversity tend to be higher in the peel, where Alphaproteobacteria and Actinobacteria dominate, while Gammaproteobacteria and Firmicutes dominate in the pulp. A statistical analysis revealed that the main characteristic features of the microbiomes of plant species originate from the peel microbiomes. Topinambur pulp displayed an interesting characteristic feature: it contained up to 108 CFUs of lactic acid bacteria, suggesting its use as a source of probiotic bacteria. We also detected Listeria sp., in topinambur pulps, however, the 16S rRNA gene fragment is unable to distinguish between pathogenic versus non-pathogenic species, so the evaluation of this potential health risk is left to a future study.

Obtaining carrot (Daucus carota L.) plants in isolated microspore cultures.

Microspores were cultured on the modified B5 liquid medium containing 2.4D (0.1 mg L(-1)), NAA (0.1 mg L(-1)), L-glutamine (500 mg L(-1), L-serine (100 mg L(-1)), and sucrose (100 g L(-1)). The developmental stages of microspores and divisions were observed. Initially, the formation of binuclear and multicellular structures was noticed. Plants regenerated in the cultures in which the tetrad stage of microsporogenesis had predominated. Embryoids were still forming 24 weeks after the cultures were set up. Six weeks after the transfer of androgenetic embryos onto the B5 regeneration medium, they were converted into complete plants. Out of 90 androgenetic plants planted in a growth chamber, 42 plants adapted to the new conditions. All of those plants proved to be diploids in cytometric analysis.

Effects of ozone treatment on Botrytis cinerea and Sclerotinia sclerotiorum in relation to horticultural product quality.

Botrytis cinerea and Sclerotinia sclerotiorum are fungal pathogens that cause the decay of many fruits and vegetables. Ozone may be used as an antimicrobial agent to control the decay. The effect of gaseous ozone on spore viability of B. cinerea and mycelial growth of B. cinerea and S. sclerotiorum were investigated. Spore viability of B. cinerea was reduced by over 99.5% (P < 0.01) and height of the aerial mycelium was reduced from 4.7 mm in the control to less than 1 mm after exposure to 450 or 600 ppb ozone for 48 h at 20 degrees C. Sporulation of B. cinerea was also substantially inhibited by ozone treatments. However, ozone had no significant effect on mycelial growth of S. sclerotiorum in vitro. Decay and quality parameters including color, chlorophyll fluorescence (CF), and ozone injury were further assessed for various horticultural commodities (apple, grape, highbush blueberry, and carrot) treated with 450 ppb of ozone for 48 h at 20 degrees C over a period of 12 d. Lesion size and height of the aerial mycelium were significantly reduced by the ozone treatment on carrots inoculated with mycelial agar plugs of B. cinerea or S. sclerotiorum. Lesion size was also reduced on treated apples inoculated with 5 x 10(6) spores/mL of B. cinerea, and decay incidence of treated grapes was reduced. The 450 ppb ozone for 48 h treatment had no significant effect on color of carrots and apples or on CF of apples and grapes. Ozone, an environmentally sound antimicrobial agent, inactivates microorganisms through oxidization and residual ozone spontaneously decomposes to nontoxic products. It may be applied to fruits and vegetables to reduce decay and extend shelf life.

The influence of several factors on the efficiency of androgenesis in carrot.

The influence of cultivar, donor plant and culture procedure on the efficiency of androgenesis was studied in carrot anther culture. Experiments were carried out on five carrot cultivars: CxC 9900 F1, Lucky B F1, HCM, Beta III and Perfekcja, which were chosen because of their high carotene contents. Two procedures of anther culture were compared: (1) incubation in darkness for two weeks, followed by exposure to continuous light and transfer onto a fresh medium of the same composition; and (2) incubation in darkness until embryos appeared, without transfer onto a fresh medium. Temperature was +27 degrees C all the time. Genotype played an important role in the process of androgenesis in carrot anther culture. The efficiency was the highest in cv. HCM - 5.6 embryos per 100 anthers. Considerable differences in the capacity for androgenesis were observed between individual donor plants. The ratio of embryos obtained per 100 anthers for cv. HCM varied from 0.0 to 48.9. The second procedure of anther culture proved to be more efficient, cheaper and less complicated.

Effects on the growth of carrots (Daucus carota L.), cabbage (Brassica oleracea var. capitata L.) and onion (Allium cepa L.) of restricting the ability of the plants to intercept resources.

The objective of this paper is to assess the size and penetration of edge effects in carrot, cabbage and onion field crops and the extent to which these edge effects are modified by the presence of aerial or soil competition between the crop rows. In all three crops, large weight differences developed between the plants in the edge rows and those in the central rows. There was no indication of plant weight fluctuating between large and small values with each successive row in from the edge, as suggested by others. In carrot and onion, edge effects were greatly reduced by the presence of either white reflective aerial partitions or soil partitions, indicating that these species competed for both light and soil resources in UK field conditions. In cabbage, the mere presence of clear aerial partitions between rows reduced edge effects and there was little effect of soil partitions. This indicates the predominance of shoot over root competition in this species. The differences between species are possibly related to the architectural flexibility of their shoots. These results suggest that, within crops, carrot and onion plants compete for light over a distance of about 20 cm in each direction and for below-ground resources over a distance of about 50 cm in each direction. For cabbage, interactions between plants appeared to be dominated by the requirement for sufficient space to deploy the shoots for efficient light interception.

Electrophysiological model of intact and processed plant tissues: cell disintegration criteria.

Frequency versus conductivity relationships of food cell system, based on impedance measurements as characterized by polarization effects of the Maxwell-Wagner type at intact membrane interfaces, are presented. The electrical properties of a biological membrane (represented as a resistor and capacitor) are responsible for the dependence of the total conductivity of the cell system on the alternating current frequency. Based on an equivalent circuit model of a single plant cell, the electrical conductivity spectrum of the cell system in intact plant tissue (potato, carrot, banana, and apple) was determined in a frequency range between 3 kHz and 50 MHz. The electrical properties of a cell system with different ratios of intact/ruptured cells could also be predicted on the basis of a description of a cell system consisting of elementary layers with regularly distributed intact and ruptured cells as well as of extracellular compartments. This simple determination of the degree of cell permeabilization (cell disintegration index, p(o)) is based upon electric conductivity changes in the cell sample. For accurate calculations of p(o), the sample conductivities before and after treatment, obtained at low- (f(l)) and high-frequency (f(h)) ranges of the so-called beta-dispersion, were used. In this study with plant cell systems, characteristic conductivities used were measured at frequencies f(l) = 3 kHz and f(h) = 12.5 MHz. The disintegration index was used to analyze the degree of cell disruption after different treatments (such as mechanical disruption, heating, freeze-thaw cycles, application of electric field pulses, and enzymatic treatment) of the plant tissues.

Characterization of chitinases able to rescue somatic embryos of the temperature-sensitive carrot variant ts 11.

To characterize the acidic endochitinase EP3, able to rescue somatic embryos of the carrot cell line ts11, the enzyme was purified from the medium of wild-type suspension cultures. Peptide sequences, deduced amino acid sequences of corresponding PCR-generated cDNA clones, serological relation and biochemical properties showed that there were at least five closely related chitinases, four of which could be identified as class IV EP3 chitinases with an apparent size of 30 kDa. Two other proteins were identified as a serologically related class I acidic chitinase (DcChitI) of 34 kDa, and a serologically unrelated 29 kDa class II acidic chitinase (DcChitII), respectively. Additional cDNA sequences, Western and Southern analysis showed the presence of a least two, but possibly more, highly homologous class IV EP3 genes in the carrot genome. Two class IV EP3 chitinases were tested and found to be able to increase the number of ts11 globular embryos formed under non-permissive conditions. One of the class IV EP3 chitinases as well as the class I chitinase DcChitI promoted the transition from globular to heart-stage ts11 embryos. The class II endochitinase and a heterologous class IV chitinase from sugar-beet were not active on ts11. This suggests that there are differences in the specificity of chitinases in terms of their effect on plant somatic embryos.